Biotechnology and Bioinformatics (1+1)

Materials required

· Xylene

· Ethanol, anhydrous denatured, histological grade (100% and 95%)

· Deionized water (dH₂O)

· Hematoxylin (optional)

· Wash Buffer: 1X TBS/0.1% Tween-20 (1X TBST)

· 10X Tris Buffered Saline (TBS) 24.2 g Trizma^® base (C₄H₁₁NO₃) and 80 g sodium chloride (NaCl) to 1 L dH₂O. Adjust pH to 7.6 with concentrated HCl.

· Antibody Diluent: TBST/5% normal goat serum

· Antigen Unmasking- Citrate ( 10 mM Sodium Citrate Buffer) or EDTA ( 1 mM EDTA)

· 3% Hydrogen Peroxide: To prepare, add 10 ml 30% H₂O₂ to 90 ml dH₂O.

· Blocking Solution

· Secondary antibody conjugate to enzyme

· Substrate solution (DAB: 3,3´-diaminobenzidine tetrahydrochloride or NBT-BCIP: 5-bromo-4-chloro-3-indolyl-phosphate/ nitro-blue tetrazolium)